Mass spectrometry reveals TRPM7-interacting proteins.

For identification of TRPM7 interacting proteins by LC-MS/MS, purified TRPM7 was resolved in a Bis-Tris polyacrylamide gel, and each gel band was subjected to in-gel reduction, alkylation, tryptic digestion, and peptide extraction with a standard protocol. Peptides were solubilized in 0.1% trifluoroacetic acid, and analyzed by Nano LC-MS/MS with Dionex Ultimate 3000 RLSC Nano System interfaced with a Velos-LTQ-Orbitrap (Thermo Fisher Scientific) or a QExactive HF (Thermo Fisher Scientific) based on instrument availability. Shown are results from samples immunopurified from LTRPC7 cells in the absence of tetracycline to keep TRPM7 expression levels low and from control cells (293T). The table shows proteins that were identified in 2 experimental replicates of samples from LTRPC7 cells but not in in control 293T cells.
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